Cell-free transcription amplification-based split-type electrochemical sensor using enzyme-linked magnetic microbeads for minimal residual leukemia detection.

Constrained by detecting techniques, patients with acute promyelocytic leukemia (APL) are often confronted with minimal residual disease (MRD) and a high risk of relapse. Thus, a pragmatic and robust method for MRD monitoring is urgently needed. Herein, a novel split-type electrochemical sensor (E-sensor) was developed by integrating nucleic acid sequence-based amplification (NASBA) with enzyme-linked magnetic microbeads (MMBs) for ultra-sensitive detection of the PML/RARα transcript.

Engineered Aptamer-Derived Fluorescent Aptasensor: the Label-Free, Single-Step, Rapid Detection of Vancomycin in Clinical Samples.

Currently, the reported vancomycin (VCM) aptamers, including the 3- (K =  9.13 × 10 m) and 4-truncated variants (K = 45.5 × 10 m), are engineered via stem truncation of the VCM parent aptamer, which inevitably compromises their affinities, thus affecting their clinical application within the VCM therapeutic window of 6.

A homogeneous electrochemiluminescence immunoassay platform based on carbon quantum dots and magnetic beads enrichment for detection of thyroglobulin in serum.

Considering the high probability of recurrence or metastasis after thyroidectomy, it is meaningful to develop a rapid, sensitive and specific method for monitoring thyrophyma-related biomarkers. In this study, a homogeneous electrochemiluminescence immunoassay (HO-ECLIA) coupled with magnetic beads (MBs)-based enrichment tactic was established for the determination of thyrophyma-related thyroglobulin (Tg). Importantly, owing to the abundant surface groups and good biocompatibility of carbon quantum dots (CQDs), the incorporation of CQDs onto the Tg antigen surface was achieved, resulting in the formation of Tg-encapsulated CQDs (CQDs-Tg), which served not only as an ECL probe but as a biorecognition element.

Precise Differentiation of Wobble-Type Allele via Ratiometric Design of a Ligase Chain Reaction-Based Electrochemical Biosensor for Genotyping of Clinical Samples.

Due to the comparable stability between the perfect-base pair and the wobble-base pair, a precise differentiation of the wobble-type allele has remained a challenge, often leading to false results. Herein, we proposed a ligase chain reaction (LCR)-based ratiometric electrochemical DNA sensor, namely, R-eLCR, for a precise typing of the wobble-type allele, in which the traditionally recognized "negative" signal of wobble-base pair-mediated amplification was fully utilized as a "positive" one and a ratiometric readout mode was employed to ameliorated the underlying potential external influence and improved its detection accuracy in the typing of the wobble-type allele. The results showed that the ratio between current of methylene blue () and current of ferrocene () was partitioned in three regions and three types of wobble-type allele were thus precisely differentiated (AA homozygote: / > 2; GG homozygote: / < 1; GA heterozygote: 1 < / < 2); the proposed R-eLCR successfully discriminated the three types of allele in nine cases of human whole blood samples, which was consistent with those of the sequencing method.

Nucleic Acid Amplification by Template-Dominated Click Chemistry for Ultrasensitive DNA/RNA Detection on an Electrochemical Readout Platform.

As an enzyme-free exponential nucleic acid amplification method, the click chemistry-mediated ligation chain reaction (ccLCR) has shown great prospects in the molecular diagnosis. However, the current optics-based ccLCR is challenged by remarkable nonspecific amplification, severely hindering its future application. This study demonstrated that the severe nonspecific amplification was generated probably due to high random collision in the high DNA probe concentration (μM level).

Electrochemical immunosensor based on AuNPs/ERGO@CNT nanocomposites by one-step electrochemical co-reduction for sensitive detection of P-glycoprotein in serum.

P-glycoprotein (P-gp), a transmembrane glycoprotein widely expressed on the surface of various cells, is highly associated with multidrug resistance (MDR) that heralds the malignant progress of disease after drug treatment. Notably, there have been reported that serum P-gp is a potential marker for assessing the progression of disease resistance. Currently, there are few methods for point-of-care serum P-gp detection.

Sequence-specific amperometric detection based on a double-probe mode and enzyme-mediated multiple signal electrocatalysis for the double-stranded DNA of PML/RARα-related fusion gene.

A new electrochemical DNA biosensor based on double-probe mode and enzyme-mediated multiple signal electrocatalysis is constructed for the highly sensitive determination of double-stranded (ds-) PML/RARα fusion gene. Through the ingenious design of two groups of detection probes, including two thiolated capture probes anchored on dual standalone detection units integrated into one customized gold electrode and four biotinylated reporter probes, hybridizing with different segments of the same target single-stranded DNA (ssDNA) simultaneously, the hybridization efficiency between the probes and target is improved by preventing the reannealing of the two separate target ssDNA. Compared with a single reporter probe, this method can dramatically increase the amount of biotin and introduce numerous streptavidin-labelled horseradish peroxidase (HRP), thereby significantly amplifying electrochemical signals with low background signals.

DNA Nanosieve-Based Regenerative Electrochemical Biosensor Utilizing Nucleic Acid Flexibility for Accurate Allele Typing in Clinical Samples.

Herein, an interface-based DNA nanosieve that has the ability to differentiate ssDNA from dsDNA has been demonstrated for the first time. The DNA nanosieve could be readily built through thiol-DNA's self-assembly on the gold electrode surface, and its cavity size was tunable by varying the concentration of thiol-DNAs. Electrochemical chronocoulometry using [Ru(NH)] as redox revealed that the average probe-to-probe separation in the 1 μM thiol-DNA-modified gold electrode was 10.

Integrated paper-based 3D platform for long-term cell culture and in situ cell viability monitoring of Alzheimer's disease cell model.

Reactive oxygen species including superoxide anion, hydrogen peroxide (HO) and hydroxyl radicals, as a conflicting class of biological metabolites in living organism, act crucial effect on Alzheimer's disease (AD). In this work, a facile integrated platform composed of a paper-based three-dimension (3D) cell culture system and an electrochemical sensor was developed for the construction of AD cell model in third dimensional structure and in situ cell viability monitoring by HO released from PC12 cells cultured on paper scaffold were divided into three groups containing control group, amyloid beta peptide 25-35 (Aβ) group and Aβ+curcumin (Aβ+cur) group, respectively. In addition, the paper-based 3D platform displayed excellent properties, such as sensitivity, selectivity, reproducibility and stability.

Ultrasensitive Detection of RNA with Single-Base Resolution by Coupling Electrochemical Sensing Strategy with Chimeric DNA Probe-Aided Ligase Chain Reaction.

Accurate and sensitive detection of single-base mutations in RNAs is of great value in basic studies of life science and medical diagnostics. However, the current available RNA detection methods are challenged by heterogeneous clinical samples in which trace RNA mutants usually existed in a large pool of normal wild sequences. Thus, there is still great need for developing the highly sensitive and highly specific methods in detecting single-base mutations of RNAs in heterogeneous clinical samples.

A novel ligase chain reaction-based electrochemical biosensing strategy for highly sensitive point mutation detection from human whole blood.

Challenged by the detection of trace amounts of mutants and disturbance from endogenous substances in clinical samples, herein, we present a novel electrochemical biosensor based on ligase chain reaction (eLCR) via the thermostable ligase with high mutation recognizing ability. The lengthened double-stranded DNAs exponentially generated via LCR were uniformly distributed on a bovine serum albumin-modified gold electrode, in which the phosphate buffer was tactfully added to remove adsorbed uninterested-probes, and thereafter the amperometry current was collected for the specific binding of streptavidin-poly-HRP and subsequent catalysis in the 3, 3', 5, 5'-tetramethylbenzidine substrate that contained hydrogen peroxide. It found that, under optimized conditions, the proposed biosensor exhibited a high selectivity of mutant targets from the 10-fold excess of co-existent wild targets within a detection limit of 0.

Sepsis progression monitoring via human serum fibronectin detection based on sandwich-type electrochemical immunosensor.

Sepsis has always been a severe clinical problem in critical care medicine due to its rather high mortality and poor prognosis. The current study reported for the first time a practical immunosensor for fibronectin (FN) detection in human serum by electrochemistry. A simple but robust sandwich-type strategy was employed without any complex design or material modifications, which exhibited a linear calibration plot over the 15.

Paper-based 3D culture device integrated with electrochemical sensor for the on-line cell viability evaluation of amyloid-beta peptide induced damage in PC12 cells.

In this communication, a paper-based 3D cell culture device integrated with electrochemical biosensor was applied to on-line monitoring of dopamine release from PC12 cell damage models induced by amyloid-beta peptide (Aβ) and cell intervene models protected by curcumin (Cur) and marrow mesenchymal stem cells (MSC) supernatant. The adhesion and proliferation of PC12 cells cultured on the paper scaffold was characterized by scanning electron microscopy and laser scanning confocal microscopy, which verify unique biocompatibility and 3D microarchitecture similar to human body microenvironment of paper substrate, so an artificial model simulating 3D microenvironment in vivo was constructed easily. The PC12 cells in paper-based devices consisted of four groups containing control group, Aβ group, Aβ+Cur group and Aβ+MSC supernatant group.

Improving quantitative control and homogeneous distribution of samples on paper-based analytical devices via drop-on-demand inkjet printing.

A standard desktop printer with multiple ink cartridges can accurately deposit a broad variety of biomaterials on microfluidic paper-based analytical devices (μPADs) which have been extensively applied to environmental monitoring and screening of food and beverage contamination. Finding ways to realize sample quantitative control by tuning the CMYK value, however, remains challenging. Herein, we studied the influence of the CMYK value on the ink volume jetted by ink cartridges.

A special issue to mark the 90th Anniversary of College of Life Sciences, Zhejiang University.

The College of Life Sciences (CLS) remains one of the most prestigious-and the oldest-colleges in Zhejiang University. This special issue, which includes 16 reviews contributed by our alumni and faculties, is dedicated to mark the 90th Anniversary of CLS. The reviews provide a glimpse of current progresses in the areas of life sciences such as biochemical processes and their association with diseases (Ding et al.

A colorimetric assay for sensitive detection of hydrogen peroxide and glucose in microfluidic paper-based analytical devices integrated with starch-iodide-gelatin system.

Microfluidic paper-based analytical devices (μPADs) for detection of hydrogen peroxide and glucose have been developed. The analytical performance of colorimetric detection using the conventional starch-iodine color reaction has been significantly improved by using gelatin as the surface modifier which retains the enzyme activity in the dry filter paper strip, improves antioxidability, as well as decreases the strong background signal. Under optimal conditions, the color intensities show a good linear relationship with glucose concentration ranging from 0.

Glomerular Filtration Rate Is a Major Predictor of Clearance of Oxcarbazepine Active Metabolite in Adult Chinese Epileptic Patients: A Population Pharmacokinetic Analysis.

Background: Oxcarbazepine (OXC) is almost completely metabolized to its10-monohydroxy derivative (MHD), which is responsible for the pharmacological effects of the drug. Several studies have described the population pharmacokinetics (PPK) of MHD in pediatric patients, but little is known about its pharmacokinetics in adult patients. In addition, no study to date has proposed a model to investigate the influence of genetic polymorphisms on MHD pharmacokinetics.

Ultrasensitive Electrochemical Biosensor Developed by Probe Lengthening for Detection of Genomic DNA in Human Serum.

As an alternative to most of the reported nucleic acid amplification-based electrochemical DNA biosensors used for detection of trace levels of genomic DNA, we herein present a novel detection concept. The proposed system involves the conversion of two short double-stranded DNAs (dsDNAs), labeled with a thiol-tag or biotin-tag, into a single integrated dsDNA containing thiol and biotin at both terminals in the presence of target DNA through ligase chain reaction (LCR) and followed by the immobilization of these integrated dsDNAs on a bovine serum albumin (BSA)-modified gold electrode surface. Owing to rapid depletion of the two short dsDNAs via LCR, the integrated dsDNAs were generated in an exponential manner so that this sensoring approach offered a limit of detection of 25 yoctomoles (15 copies in 50 μL sample volumes), a high discrimination of single-base mismatch and a wide linear concentration range (across 6 orders of magnitude) for target DNA.

2'-Fluoro ribonucleic acid modified DNA dual-probe sensing strategy for enzyme-amplified electrochemical detection of double-strand DNA of PML/RARα related fusion gene.

In the study, a novel sensing strategy based on dual-probe mode, which involved two groups of 2'-fluoro ribonucleic acid (2'-F RNA) modified probes, was designed for the detection of synthetic target double-strand DNA (dsDNA) of PML/RARα fusion genes in APL. And each pair of probes contained a thiolated capture probe (C1 or C2) immobilized on one of electrode surfaces in the dual-channel electrochemical biosensor and a biotinylated reporter probe (R1 or R2). The two groups of 2'-F RNA modified probes were separately complementary with the corresponding strand (Sa or Sb) from target dsDNA in order to prevent renaturation of target dsDNA.

Adropin deficiency worsens HFD-induced metabolic defects.

The limited efficacy of current treatment methods and increased type 2 diabetes mellitus (T2DM) incidence constitute an incentive for investigating how metabolic homeostasis is maintained, to improve treatment efficacy and identify novel treatment methods. We analyzed a three-generation family of Chinese origin with the common feature of T2DM attacks and fatty pancreas (FP), alongside 19 unrelated patients with FP and 58 cases with T2DM for genetic variations in Enho, serum adropin, and relative T amounts. Functional studies with adropin knockout (AdrKO) in C57BL/6J mice were also performed.

CALCB splice region pathogenic variants leading to plasma cell neurotropic enrichment in type 1 autoimmune pancreatitis.

Recently, we have demonstrated that PRSS1 mutations cause ectopic trypsinogen activation and thereby result in type 1 autoimmune pancreatitis (AIP). However, the molecules involved in inducing obliterative vasculitis and perineural inflammation in the pancreas are not well-described. The present study applied whole-exome sequencing (WES) to determine the underlying etiology and revealed novel missense splice region variants, CALCB c.